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Kwan Lab
Добавлен 14 янв 2016
Видео
Expression and purification of His-tagged proteins from E. coli
Просмотров 88 тыс.3 года назад
Expression and purification of His-tagged proteins from E. coli
[BIOL 515] Preparation for Module 2 - RNA Extraction from Yeast
Просмотров 3,8 тыс.3 года назад
[BIOL 515] Preparation for Module 2 - RNA Extraction from Yeast
[BIOL 515] Preparation for Modules 2 & 3 - Culturing and harvesting yeast cells
Просмотров 3663 года назад
[BIOL 515] Preparation for Modules 2 & 3 - Culturing and harvesting yeast cells
[Biol 515] Module 1 - Part A: RNA Extraction from E. coli
Просмотров 2,1 тыс.3 года назад
[Biol 515] Module 1 - Part A: RNA Extraction from E. coli
[Biol 515] Module 0 - Part B: Running DNA samples on agarose gel
Просмотров 2905 лет назад
[Biol 515] Module 0 - Part B: Running DNA samples on agarose gel
[Biol 515] Module 0 - Part A: Use of Micropipettors
Просмотров 3355 лет назад
[Biol 515] Module 0 - Part A: Use of Micropipettors
Thanks a lot :)
Can we resuse this columns.. Or we need to add agarose colum each time?
Awesome video, but it's criminal that you didn't show how the gel looks like after coomassie destaining
Which protein you are expressing and purifying
U have used dnase before sonication.. does the addition of dnase before sonication impacts lysis differently or it does not affect the lysis
On original is omeletu formage
It extends 3' end 😂
😂😂My type of content. I'm home now 😅
Thank you !!!
Can you share some references you use to do in this video? Thank you so much
Thanks for the video. As a first-year PhD student, I needed this video to get my feet on the ground. Thanks a lot
All Biochemistry Master's Students looking to understand the method of protein expression for your project hands up ✋🏽
holler 🖐
Hi where is your lab I have a few questions
School crushes be like
Please tell me how you made your elution buffer, everytime I add imidazole the pH rises above whats needed. And adjusting it with acid is not possible because of possible interference.
It is always better to add lysozyme AFTER resuspending the pellet.
🤠
lmao 5' to 3' always!
Great performance, great explanation. Thank you.
Quality content
first of all - thanks, it really helped me to understand several steps of this procedure better! secondly, it's the best unintentional asmr i've heard in my entire life, please, continue
So helpful, thanks a lot!!
love this video thank you!!
10:40 for Day 4
I'm glad that I found your channel. It's really a nice and detailed Protein extraction video on RUclips that's really helpful for beginners like me 😃
How do you determine what gradient of SDS-PAGE gel to use since there are multiple products on the market that range from 4%-12, or 4%-20% gradient?
Really good and well explained 👍
Hi Great Job Can I have this protocol in written form So i cannot miss any point. It would be highly appreciated
I appreciated the level of detail you described here, thanks. Using your technique, about how long does it take you to grow and purify a 1L culture?
What is the success/trial ratio of this process, if i follow the process exactly, will i be able to express the protein or does it takes few trials
Great video! Great technique. Thank you.
Hi, thank you very much for your excellent job! And I have a question, at 1'43'', what is the work frequency of the machine? Thank you!
whats the name of the spectrophotometer machine you were using?
Hi, this video was great!! Maybe you can do additional videos on SDS-page interpretation (maybe with different proteins and conditions).
LB should be pH'd to 7.
I want to see how the running gel looks like.
thank you the videos ,i am doing sds page but my run wont start even after adding running buffer to the mark .i use the same mini gel tank like yours ,though it leaks ,could it be the issue ?help me from this confusion please
Use fresh running buffer, ensure you have the electrodes the correct way, ensure no leaks, make sure gel is entered the right way, remove the white tape at the bottom of the pre-case gel if you are using those
Thank You so much sir
Outstanding sir
Hello! My name is Maria and I am a PhD student working on a project that works to collate various biological techniques for early career researchers in the lab. I really loved your tutorial video for protein purification, and was wondering whether we could get in touch to discuss it further and other similar protocols. Please let me know I'd love to hear back from you!
Thank you very much, this is great for teaching with limited lab.
Aabsolute master! Thanks my dude - this helped a lot! For the viewer's sake you might include on-screen stats for the reagents used. Thanks again! :D
I liked the cotton idea :) on gel staining part of the protocol. Useful indeed
Do you have a protocol please
Please continue this, it's definitely a channel worth subscribing
I do protein expression with 6 liters of culture. After centrifugation of the lysate, the supernatant is passed onto the Ni-NTA agarose column. It takes several days for the supernatant from such a large culture to pass through the column or immediately by syphoning the supernatant out of the column by applying a vacuum to the base of the column.
Use a 5 mL HiTrap with an FPLC system
please write the name of manufacture Ni-nickel resin
Thank you